1) The OD of an E.coli culture is 0.46. You transfer 50µl of this culture into 4

Q: 1) The OD of an E.coli culture is 0.46. You transfer 50µl of this culture into 4

Based on the given data, The OD of an E.coli culture is 0.46, so the culture has 5×107 cells/ml. So, the pattern of dilution is: Based on the above dilution, the LA plate must have 125 colonies, but not 12 colonies. It is because at the last dilation 1 ml has 5000 cells, so 25µl has 125 cells. Therefore, each cell is generated into a colony the plate should have 125 colonies.

ID: 52537 • Letter: 1

Question

1)The OD of an E.coli culture is 0.46. You transfer 50µl of this culture into 4.95ml of medium. You repeat this same dilution one more time (This is your dilution 2). You transfer 25 µl of your dilution 2 onto an LA plate. After 24 hours of incubation, you observe 12 colonies on this plate. A) Did you expect this result? If not, name at least three reasons for this result giving a detailed explanation for each reason. B) What would you do to prove your theory and why?

Note: E. coli at OD595 at 1.0 is 10^8 cells/ml.

Explanation / Answer

Based on the given data,

The OD of an E.coli culture is 0.46, so the culture has 5×107 cells/ml. So, the pattern of dilution is:

Based on the above dilution, the LA plate must have 125 colonies, but not 12 colonies. It is because at the last dilation 1 ml has 5000 cells, so 25µl has 125 cells. Therefore, each cell is generated into a colony the plate should have 125 colonies.

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1) The OD of an E.coli culture is 0.46. You transfer 50µl of this culture into 4

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